The efficiency of shotgun proteomic analysis is dependent on the reproducibility of the peptide cleavage process during sample preparation. To generate rapid and useful metrics for peptide cleavage efficiency, as in enzymatic or chemical cleavage, SPACEPro was developed to evaluate efficiency and reproducibility of protein cleavage in peptide samples following data-dependent analysis by mass spectrometry. SPACEPro analyzes samples at the peptide-spectrum match (PSM), peptide, and protein levels to provide a comprehensive representation of the overall sample processing to peptides. All output is provided in human-readable text and JSON files that can be further processed to assess the cleavage efficiency on proteins within the sample. SPACEPro provides a snapshot of the protein cleavage efficiency through very minimal effort so that the user is informed of the quality of the sample processing efficiency and can accordingly develop protocols to improve the initial sample preparation for subsequent analyses.
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http://dx.doi.org/10.1021/acs.jproteome.0c00928 | DOI Listing |
Bioinform Adv
December 2024
Digital Technologies Research Center, National Research Council Canada, Ottawa, ON, K1A 0R6, Canada.
Motivation: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 system is a ground-breaking genome editing tool, which has revolutionized cell and gene therapies. One of the essential components involved in this system that ensures its success is the design of an optimal single-guide RNA (sgRNA) with high on-target cleavage efficiency and low off-target effects. This is challenging as many conditions need to be considered, and empirically testing every design is time-consuming and costly.
View Article and Find Full Text PDFBMC Biotechnol
January 2025
National Food Institute, Technical University of Denmark, Kgs. Lyngby, 2800, Denmark.
Background: With the growing interest in applying fermentation to seaweed biomasses, there is a need for fast and efficient selection of microbial strains that have the ability to 1) acidify quickly, 2) utilize seaweed constituents and c) exhibit some proteolytic activity. The present study aims to provide a fast methodology to screen large bacterial collections for potential applications in optimized seaweed fermentations, as well as investigate and assess the performance of a selected bacterial collection of the National Food Institute Culture Collection (NFICC) in seaweed fermentation. This approach is directed toward high-throughput (HT) methodologies, employing microwell assays for different phenotypical characteristics of lactic acid bacteria isolated from different sources.
View Article and Find Full Text PDFJ Reprod Dev
December 2024
Division of Dairy Cattle Feeding and Breeding Research, Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Ibaraki, 305-0901, Japan.
To produce an embryo, a high conception rate must be complied along with four evaluation criteria based on the timing of early cleavage and proper embryo morphology (hereafter, these blastocysts will be referred to as "four-criteria-compliant blastocysts"). Therefore, it is necessary to construct a culture system for high efficiency production of embryos meeting these four criteria. Non-essential amino acids (NEAAs) are widely used for the culture of bovine embryos fertilized in vitro; however, the necessity and optimal concentration of individual NEAA must be verified to produce four-criteria-compliant blastocysts.
View Article and Find Full Text PDFBiol Reprod
January 2025
School of Animal Sciences, Virginia Tech, Blacksburg, Virginia, USA.
This work explored whether bovine embryo development relies on signaling from the interleukin-6 (IL6) cytokine family. This was accomplished by interrupting IL6 signal transducer (IL6ST), the common beta-subunit receptor used by the IL6 family. One series of studies cultured in vitro-produced (IVP) embryos with SC144, a pharmacological IL6ST inhibitor.
View Article and Find Full Text PDFVet Microbiol
December 2024
National Key Laboratory of Veterinary Public Health and Safety, Key Laboratory of Animal Epidemiology of the Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
Pseudorabies virus (PRV) poses a significant threat to the global swine breeding industry and public health, but how the virus transverses the host defense systems for efficient viral replication and pathogenesis remains unclear. Here, we report that PRV could inhibit the unfolded protein response (UPR), a critical component of host innate immunity against viral infection, to promote virus replication during the late infection stages. PERK was shown phosphorylated and active in PRV-infected cells, but the subsequent events were suppressed post virus infection, such as eIF2α phosphorylation, ATF4 expression, and the formation of stress granules (SGs).
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